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Human 4-Hydroxynonenal(HNE) ELISA Kit
ELK8372
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48 T $320.00
96 T $458.00
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OverviewPublications

Product name: Human 4-Hydroxynonenal(HNE) ELISA Kit
Reactivity: Human
Alternative Names: 4-HNE; HNE; 4-hydroxy-2-nonenal; 4-Hydroxynonenal
Assay Type: Competitive Inhibition
Sensitivity: 19.1 pg/mL
Standard: 2500 pg/mL
Detection Range: 39.07-2500 pg/mL
Sample Type: Serum, plasma, tissue homogenates
Assay Length: 2h
Research Area: Signal transduction;Metabolic pathway;Infection immunity;Hormone metabolism;Dermatology;
Test principle: This assay employs the competitive inhibition enzyme immunoassay technique. The microtiter plate provided in this kit has been pre-coated with Human 4-HNE protein. Standards or samples are added to the appropriate microtiter plate wells then with a biotin-conjugated antibody specific to Human 4-HNE. Next, Avidin conjugated to Horseradish Peroxidase (HRP) is added to each microplate well and incubated. After TMB substrate solution is added. The enzyme-substrate reaction is terminated by the addition of sulphuric acid solution and the color change is measured spectrophotometrically at a wavelength of 450nm ± 10nm. The concentration of Human 4-HNE in the samples is then determined by comparing the OD of the samples to the standard curve.

Standard curve

Concentration (pg/mL) OD Corrected OD
2500.00 0.232
1250.00 0.336
625.00 0.583
312.50 0.993
156.25 1.225
78.13 1.596
39.07 1.877
0.00 2.325

Precision

Intra-assay Precision (Precision within an assay)：CV%<8%

Three samples of known concentration were tested twenty times on one plate to assess intra-assay precision.

Inter-assay Precision (Precision between assays)：CV%<10%

Three samples of known concentration were tested in forty separate assays to assess inter-assay precision.

Recovery

Matrices listed below were spiked with certain level of recombinant 4-HNE and the recovery rates were calculated by comparing the measured value to the expected amount of 4-HNE in samples.
Matrix Recovery range Average
serum(n=5) 85-97% 91%
EDTA plasma(n=5) 83-95% 89%
Heparin plasma(n=5) 80-92% 101%

Linearity

The linearity of the kit was assayed by testing samples spiked with appropriate concentration of 4-HNE and their serial dilutions. The results were demonstrated by the percentage of calculated concentration to the expected.
Matrix 1:2 1:4 1:8 1:16
serum(n=5) 89-97% 85-94% 92-102% 81-93%
EDTA plasma(n=5) 88-102% 87-102% 85-96% 87-101%
Heparin plasma(n=5) 87-95% 85-96% 92-102% 81-92%